scholarly journals Exploitation of bacterial N -linked glycosylation to develop a novel recombinant glycoconjugate vaccine against Francisella tularensis

Open Biology ◽  
2013 ◽  
Vol 3 (5) ◽  
pp. 130002 ◽  
Author(s):  
Jon Cuccui ◽  
Rebecca M. Thomas ◽  
Madeleine G. Moule ◽  
Riccardo V. D'Elia ◽  
Thomas R. Laws ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Campylobacter Jejuni ◽  
Francisella Tularensis ◽  
Single Step ◽  
Carrier Protein ◽  
Purification Process ◽  
Bacterial Protein ◽  
Time To Death ◽  
Glycoconjugate Vaccines ◽  
Glycoconjugate Vaccine

Glycoconjugate-based vaccines have proved to be effective at producing long-lasting protection against numerous pathogens. Here, we describe the application of bacterial protein glycan coupling technology (PGCT) to generate a novel recombinant glycoconjugate vaccine . We demonstrate the conjugation of the Francisella tularensis O-antigen to the Pseudomonas aeruginosa carrier protein exotoxin A using the Campylobacter jejuni PglB oligosaccharyltransferase . The resultant recombinant F. tularensis glycoconjugate vaccine is expressed in Escherichia coli where yields of 3 mg l −1 of culture were routinely produced in a single-step purification process. Vaccination of BALB/c mice with the purified glycoconjugate boosted IgG levels and significantly increased the time to death upon subsequent challenge with F. tularensis subsp. holarctica . PGCT allows different polysaccharide and protein combinations to be produced recombinantly and could be easily applicable for the production of diverse glycoconjugate vaccines.

scholarly journals Synthetic carbohydrate-based vaccines: challenges and opportunities

2020 ◽  
Vol 27 (1) ◽  
Author(s):  
Ravinder Mettu ◽  
Chiang-Yun Chen ◽  
Chung-Yi Wu
Keyword(s):  
Bacterial Infections ◽  
Synthetic Methods ◽  
Carrier Protein ◽  
Capsular Polysaccharides ◽  
Well Control ◽  
Bacterial Fermentation ◽  
Challenges And Opportunities ◽  
Glycoconjugate Vaccines ◽  
Glycoconjugate Vaccine ◽  
Synthetic Carbohydrate

AbstractGlycoconjugate vaccines based on bacterial capsular polysaccharides (CPS) have been extremely successful in preventing bacterial infections. The glycan antigens for the preparation of CPS based glycoconjugate vaccines are mainly obtained from bacterial fermentation, the quality and length of glycans are always inconsistent. Such kind of situation make the CMC of glycoconjugate vaccines are difficult to well control. Thanks to the advantage of synthetic methods for carbohydrates syntheses. The well controlled glycan antigens are more easily to obtain, and them are conjugated to carrier protein to from the so-call homogeneous fully synthetic glycoconjugate vaccines. Several fully glycoconjugate vaccines are in different phases of clinical trial for bacteria or cancers. The review will introduce the recent development of fully synthetic glycoconjugate vaccine.

scholarly journals Mutagenesis and immunological evaluation of group A streptococcal C5a peptidase as an antigen for vaccine development and as a carrier protein for glycoconjugate vaccine design

RSC Advances ◽  
2017 ◽  
Vol 7 (67) ◽  
pp. 42056-42063 ◽  
Author(s):  
Hui Li ◽  
Subo Wang ◽  
Yisheng Zhao ◽  
Zonggang Chen ◽  
Guofeng Gu ◽  
...  
Keyword(s):  
Vaccine Development ◽  
Vaccine Design ◽  
Carrier Protein ◽  
Group A ◽  
Application Potential ◽  
Glycoconjugate Vaccines ◽  
Glycoconjugate Vaccine ◽  
Immunological Evaluation

A non-enzymatic recombinant ScpA mutant (H193A) was prepared and investigated to probe its application potential in the development of GAS vaccines and as a carrier protein of glycoconjugate vaccines.

scholarly journals The Stability of Acyl Carrier Protein in Escherichia coli

1973 ◽  
Vol 248 (12) ◽  
pp. 4461-4466
Author(s):  
Gary L. Powell ◽  
Michael Bauza ◽  
Allan R. Larrabee
Keyword(s):  
Escherichia Coli ◽  
Acyl Carrier Protein ◽  
Carrier Protein ◽  
The Stability

scholarly journals Mutants of Escherichia coli with Temperature-sensitive Malonyl Coenzyme A-Acyl Carrier Protein Transacylase

1974 ◽  
Vol 249 (23) ◽  
pp. 7468-7475
Author(s):  
Mark E. Harder ◽  
Ruth C. Ladenson ◽  
Steven D. Schimmel ◽  
David F. Silbert
Keyword(s):  
Escherichia Coli ◽  
Coenzyme A ◽  
Acyl Carrier Protein ◽  
Temperature Sensitive ◽  
Carrier Protein

scholarly journals Morphology of proteoliposomes reconstituted with purified lac carrier protein from Escherichia coli.

1984 ◽  
Vol 259 (24) ◽  
pp. 15579-15586 ◽  
Author(s):  
M J Costello ◽  
P Viitanen ◽  
N Carrasco ◽  
D L Foster ◽  
H R Kaback
Keyword(s):  
Escherichia Coli ◽  
Carrier Protein

scholarly journals Random and combinatorial mutagenesis for improved total production of secretory target protein in Escherichia coli

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
David Gonzalez-Perez ◽  
James Ratcliffe ◽  
Shu Khan Tan ◽  
Mary Chen May Wong ◽  
Yi Pei Yee ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Protein Secretion ◽  
Protein Production ◽  
Secretory Protein ◽  
Target Protein ◽  
Carrier Protein ◽  
Signal Peptides ◽  
Carrier Proteins ◽  
E Coli ◽  
Combinatorial Mutagenesis

AbstractSignal peptides and secretory carrier proteins are commonly used to secrete heterologous recombinant protein in Gram-negative bacteria. The Escherichia coli osmotically-inducible protein Y (OsmY) is a carrier protein that secretes a target protein extracellularly, and we have previously applied it in the Bacterial Extracellular Protein Secretion System (BENNY) to accelerate directed evolution. In this study, we reported the first application of random and combinatorial mutagenesis on a carrier protein to enhance total secretory target protein production. After one round of random mutagenesis followed by combining the mutations found, OsmY(M3) (L6P, V43A, S154R, V191E) was identified as the best carrier protein. OsmY(M3) produced 3.1 ± 0.3 fold and 2.9 ± 0.8 fold more secretory Tfu0937 β-glucosidase than its wildtype counterpart in E. coli strains BL21(DE3) and C41(DE3), respectively. OsmY(M3) also produced more secretory Tfu0937 at different cultivation temperatures (37 °C, 30 °C and 25 °C) compared to the wildtype. Subcellular fractionation of the expressed protein confirmed the essential role of OsmY in protein secretion. Up to 80.8 ± 12.2% of total soluble protein was secreted after 15 h of cultivation. When fused to a red fluorescent protein or a lipase from Bacillus subtillis, OsmY(M3) also produced more secretory protein compared to the wildtype. In this study, OsmY(M3) variant improved the extracellular production of three proteins originating from diverse organisms and with diverse properties, clearly demonstrating its wide-ranging applications. The use of random and combinatorial mutagenesis on the carrier protein demonstrated in this work can also be further extended to evolve other signal peptides or carrier proteins for secretory protein production in E. coli.

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